IL-17A neutralizing antibody regulates monosodium urate crystal-induced gouty inflammation. (September 2019)

• Record Type: Journal Article
• Authors: Raucci, Federica; Iqbal, Asif J.; Saviano, Anella; Minosi, Paola; Piccolo, Marialuisa; Irace, Carlo; Caso, Francesco; Scarpa, Raffaele; Pieretti, Stefano; Mascolo, Nicola; Maione, Francesco
• Is Part Of: Pharmacological research. Volume 147(2019)
• Publication Date: 2019-09
• Subjects: Ab antibody -- BLC B lymphocyte chemoattractant -- BSA bovine serum albumin -- Cyto-chemokines cytokines and chemokines -- COXs cyclogeneses -- C5a complement component 5a -- DMSO dimethylsulfoxide -- EDTA ethylenediamine tetraacetic acid -- EGTA ethylene glycol tetraacetic acid -- FACS fluorescence activated cell sorting -- FBS fetal bovine serum -- AGA gouty arthritis -- H&E haematoxylin and eosin -- i.a intra-articular -- IL interleukin -- IP-10 inducible protein 10 -- KC keratinocyte chemoattractant -- MCP-1 monocyte chemoattractant protein-1 -- mPGEs microsomal prostaglandin synthetase -- MHC major histocompatibility complex -- MIP macrophage inflammatory proteins -- MPO myeloperoxidase -- MSU monosodium urate -- PBS phosphate buffered saline -- PGE2 prostaglandin E2 -- PMSF phenylmethansulfonyl fluoride -- RA rheumatoid arthritis -- RANTES regulated on activation normal T expressed and secreted -- RT room temperature -- SDF-1 stromal cell-derived factor-1 -- TIMP-1 metallopeptidase inhibitor-1 -- TNF-α tumor necrosis factor-alfa -- TREM-1 triggering receptor expressed on myeloid cells-1; Ethanol (CID:702) -- PGE2 (CID:5280360) -- Phosphate-buffered saline (CID: 24978514) -- Sodium chloride (CID:5234) -- Sodium urate (CID: 23697816); Gout -- IL-17A -- Inflammation -- Monosodium urate crystal -- Neutralizing antibody; Pharmacology -- Periodicals; Pharmacology -- Periodicals; Research -- Periodicals; Médicaments -- Recherche -- Périodiques; Pharmacologie -- Périodiques; 615.105
• DOI: 10.1016/j.phrs.2019.104351
• Languages: English
• ISSNs: 1043-6618

Abstract

Graphical abstract: Abstract: Gout is a paradigm of acute, self-limiting inflammation caused by the deposition of monosodium urate (MSU) crystals within intra-and/or peri-articular areas, leading to excruciating pain, joint swelling and stiffness. The infiltration of leukocytes drives the inflammatory response and remains an attractive target for therapeutic intervention. In this context, emerging evidence supports the view that systemic differentiation of Th17 cells and their in situ infiltration as one of the potential mechanisms by which these cells, and their main product IL-17, causes damage to target tissues. To test if IL-17 was having a detrimental role in gouty onset and progression we targeted this cytokine, using a neutralizing antibody strategy, in an experimental model of gout. Joint inflammation was induced in CD-1 mice by the intra-articular (i.a.) administration of MSU crystals (200 μg/20 μl). Animals from IL-17Ab-treated groups received 1, 3 and 10 μg (i.a.) in 20 μl of neutralizing antibody after MSU crystals administration. Thereafter, joints were scored macroscopically, and knee joint oedema determined with a caliper. Histological analysis, myeloperoxidase assay and western blots analysis for COX-2/mPGEs-1/IL-17R pathway were conducted at 18 h (peak of inflammation) to evaluate leukocytes infiltration and activation, followed by the analysis, in situ, of pro/anti-inflammatory cytokines and chemokines. Flow cytometry was also used to evaluate the modulation of infiltrated inflammatory monocytes and systemic Th17 and Treg profile. Treatment with IL-17Ab revealed a dose-dependent reduction of joint inflammation scores with maximal inhibition at 10 μg. The neutralizing antibody was also able to significantly reduce leukocytes infiltration and MPO activity as well the expression of JE, IL-1α, IL-1β, IL-16, IL-17, C5a, BLC and, with a less extent IP-10, Rantes, KC, TIMP-1, SDF-1 and metalloproteinases in inflamed tissues. Biochemical analysis also revealed that IL-17Ab treatment modulated COX-2/mPGEs-1 pathway (and related PGE2 production) without interfering with IL-17R expression. Furthermore, flow cytometry analysis highlighted a selective modulation of infiltrating inflammatory monocytes (B220 - /GR1 hi -F480 hi /CD115 + ) and circulating Th17, but not Treg, cells after IL-17Ab treatment. Collectively the results of this study report for the first time, that i.a. injection of MSU crystals stimulates in vivo production of Th17 cells and Th17-related inflammatory cyto-chemokines. In addition, we have demonstrated that the administration of a neutralizing antibody against IL-17 attenuates joint symptoms, swelling and leukocytes infiltration to the inflamed tissue, possibly providing a new strategy for the treatment of gouty inflammation and/or arthritis.